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Escherichia coli K-12 substr. MG1655 Polypeptide: vitamin B12 ABC transporter - ATP binding subunit



Gene: btuD Accession Numbers: EG10128 (EcoCyc), b1709, ECK1707

Regulation Summary Diagram: ?

Component of:
vitamin B12 ABC transporter (extended summary available)
vitamin B12 transport system

Summary:
BtuD is the ATP-binding component of the BtuCD, an ABC-type vitamin B12 uptake system.

A Tn10 insertion in the btuCED promoter region restores colony-forming ability to an rne mutant. The suppression phenotype is reversed by overexpression of btuC, E or D [Tamura12].

Gene Citations: [Rioux89]

Locations: inner membrane, cytosol

Map Position: [1,790,833 <- 1,791,582] (38.6 centisomes)
Length: 750 bp / 249 aa

Molecular Weight of Polypeptide: 27.081 kD (from nucleotide sequence), 29.0 kD (experimental) [deVeaux86 ]

Unification Links: ASAP:ABE-0005707 , CGSC:18499 , DIP:DIP-9234N , EchoBASE:EB0126 , EcoGene:EG10128 , EcoliWiki:b1709 , Mint:MINT-6803741 , ModBase:P06611 , OU-Microarray:b1709 , PortEco:btuD , PR:PRO_000022236 , Protein Model Portal:P06611 , RefSeq:NP_416224 , RegulonDB:EG10128 , SMR:P06611 , String:511145.b1709 , UniProt:P06611

Relationship Links: InterPro:IN-FAMILY:IPR003439 , InterPro:IN-FAMILY:IPR003593 , InterPro:IN-FAMILY:IPR017871 , InterPro:IN-FAMILY:IPR023693 , InterPro:IN-FAMILY:IPR027417 , PDB:Structure:1L7V , PDB:Structure:2QI9 , PDB:Structure:4DBL , PDB:Structure:4FI3 , Pfam:IN-FAMILY:PF00005 , Prosite:IN-FAMILY:PS00211 , Prosite:IN-FAMILY:PS50893 , Smart:IN-FAMILY:SM00382

In Paralogous Gene Group: 23 (75 members)

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0015889 - cobalamin transport Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, deVeaux86]
GO:0035461 - vitamin transmembrane transport Inferred by computational analysis Inferred from experiment [Borths05, GOA06, GOA01]
GO:0006200 - ATP catabolic process Inferred by computational analysis [GOA01a]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11]
GO:0008152 - metabolic process Inferred by computational analysis [UniProtGOA11]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Korkhov12, Goetz09, Locher02]
GO:0015420 - cobalamin-transporting ATPase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, Borths05]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11, GOA06, GOA01a, Friedrich86]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProtGOA11]
GO:0016887 - ATPase activity Inferred by computational analysis [GOA01a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment [deVeaux86]
GO:0019898 - extrinsic component of membrane Inferred from experiment [deVeaux86]
GO:0031234 - extrinsic component of cytoplasmic side of plasma membrane Inferred from experiment [deVeaux86]
GO:0043190 - ATP-binding cassette (ABC) transporter complex Inferred from experiment Inferred by computational analysis [GOA06, Borths05]
GO:0005886 - plasma membrane Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, DiazMejia09]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11]

MultiFun Terms: metabolism biosynthesis of building blocks cofactors, small molecule carriers cobalamin
transport Channel-type Transporters Pyrophosphate Bond (ATP; GTP; P2) Hydrolysis-driven Active Transporters The ATP-binding Cassette (ABC) Superfamily + ABC-type Uptake Permeases ABC superfamily ATP binding cytoplasmic component

Essentiality data for btuD knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Subunit of: vitamin B12 ABC transporter

Subunit composition of vitamin B12 ABC transporter = [BtuD]2[BtuC]2[BtuF]
         vitamin B12 ABC transporter - ATP binding subunit = BtuD (summary available)
         vitamin B12 ABC transporter - membrane subunit = BtuC (summary available)
         vitamin B12 ABC transporter - periplasmic binding protein = BtuF (summary available)

Component of: vitamin B12 transport system

Summary:
BtuCD-F is a vitamin B12 transport system that is a member of the ATP-Binding Cassette (ABC) Superfamily of transporters [Wu95]. Based on sequence similarity, BtuD is the ATP-binding component, BtuC is the integral membrane component, and BtuF is the periplasmic substrate-binding component of the ABC transporter [Friedrich86, Borths02] . Transposon insertions in the btuC and btuD regions conferred a deficiency in vitamin B12 utilization and transport which could be complemented by plasmids carrying the corresponding functional gene [deVeaux86].

Crystal structures of the Escherichia coli BtuCD protein [Locher02] and BtuCD-F [Hvorup07] have been resolved to 3.2 Å and 2.6 Å resolution, respectively. A crytal structure of disulphide stabilized BtuCD-F in complex with the non-hydrolysable ATP analogue AMP-PNP has been resolved at 3.5 Ås [Korkhov12a]. Two BtuC subunits are in close contact and provide 20 transmembrane helices that form a translocation pathway wide enough to accomodate a vitamin B12 molecule. A short helical region in each BtuC subunit forms a cytoplasmic loop (the L-loop) which interacts with the BtuD subunits that constitute the nucleotide binding domain. The membrane spanning BtuC subunits form an enclosed substrate translocation chamber and trapping of radiolabelled cyanocobalamin in the chamber has been demonstrated [Korkhov12a]. BtuF forms a stable complex with BtuCD [Borths02, Lewinson10].

BtuCD-F is a type II ABC transporter and its transport mechanism is distinct from the well characterised type I ABC transporters such as the maltose ABC tranporter and from ABC efflux transporters [Lewinson10, Korkhov12a]. ATP binding is essential for loading vitamin B12 into the translocation chamber and hydrolysis of ATP is required for release of substrate into the cytoplasm [Korkhov12a]. In vitro purified BtuCD and BtuCD-F do not bind vitamin B12. Invitro purified BtuF associates tightly with vitamin B12 [Lewinson10]. Vitamin B12 accelerates BtuCD-F complex dissociation approximately 107 fold [Lewinson10].

The btuE gene, which resides within the btu operon does not appear to be required for vitamin B12 transport. btuE mutants have little effect on vitamin B12 binding and transport and do not affect the utilization of vitamin B12 or other cobalamins for methionine biosynthesis [Rioux89].

Citations: [Locher04, Goetz09, Oloo04, Borths05, Ivetac07, Kandt10, Joseph11, Cho13]

Locations: inner membrane

Relationship Links: PDB:Structure:1L7V , PDB:Structure:2QI9 , PDB:Structure:4DBL , PDB:Structure:4FI3

GO Terms:

Biological Process: GO:0015889 - cobalamin transport Inferred from experiment [deVeaux86, Borths05]
Molecular Function: GO:0015420 - cobalamin-transporting ATPase activity Inferred from experiment [Borths05]
Cellular Component: GO:0005887 - integral component of plasma membrane Inferred from experiment Inferred by computational analysis [Friedrich86, Locher02]
GO:0043190 - ATP-binding cassette (ABC) transporter complex Inferred by computational analysis Inferred from experiment [Borths05, Friedrich86]

Credits:
Last-Curated ? 11-Oct-2012 by Mackie A , Macquarie University


Enzymatic reaction of: Vitamin B12 ABC transporter (vitamin B12 ABC transporter)

EC Number: 3.6.3.33

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
ATP
53.0, 77.0
0.3, 1.39
[Di11, BRENDA14]


Subunit of: vitamin B12 transport system

Subunit composition of vitamin B12 transport system = [(BtuB)([TonB][ExbB][ExbD])][(BtuD)2(BtuC)2(BtuF)]
         vitamin B12 outer membrane transport complex = (BtuB)([TonB][ExbB][ExbD])
                 vitamin B12 / E colicin / bacteriophage BF23 outer membrane porin BtuB = BtuB (extended summary available)
                 TonB energy transducing system = (TonB)(ExbB)(ExbD) (extended summary available)
                         TonB energy transducing system - TonB subunit = TonB (extended summary available)
                         TonB energy transducing system - ExbB subunit = ExbB (summary available)
                         tonB energy transducing system - ExbD subunit = ExbD (summary available)
         vitamin B12 ABC transporter = (BtuD)2(BtuC)2(BtuF) (extended summary available)
                 vitamin B12 ABC transporter - ATP binding subunit = BtuD (summary available)
                 vitamin B12 ABC transporter - membrane subunit = BtuC (summary available)
                 vitamin B12 ABC transporter - periplasmic binding protein = BtuF (summary available)


Enzymatic reaction of: cobinamide transport (vitamin B12 transport system)


Enzymatic reaction of: cob(I)alamin transport (vitamin B12 transport system)

Alternative Products for cob(I)alamin: coenzyme B12 [Bradbeer78 ]


Sequence Features

Feature Class Location Citations Comment
Conserved-Region 1 -> 233
[UniProt09]
UniProt: ABC transporter;
Nucleotide-Phosphate-Binding-Region 33 -> 40
[UniProt10]
UniProt: ATP; Non-Experimental Qualifier: potential;


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b1709 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10128; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Borths02: Borths EL, Locher KP, Lee AT, Rees DC (2002). "The structure of Escherichia coli BtuF and binding to its cognate ATP binding cassette transporter." Proc Natl Acad Sci U S A 2002;99(26);16642-7. PMID: 12475936

Borths05: Borths EL, Poolman B, Hvorup RN, Locher KP, Rees DC (2005). "In vitro functional characterization of BtuCD-F, the Escherichia coli ABC transporter for vitamin B12 uptake." Biochemistry 44(49);16301-9. PMID: 16331991

Bradbeer78: Bradbeer C, Kenley JS, Di Masi DR, Leighton M (1978). "Transport of vitamin B12 in Escherichia coli. Corrinoid specificities of the periplasmic B12-binding protein and of energy-dependent B12 transport." J Biol Chem 253(5);1347-52. PMID: 342526

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Cho13: Cho YW, Kim SY, Kwon IC, Kim IS (2013). "Complex adaptive therapeutic strategy (CATS) for cancer." J Control Release 175C;43-47. PMID: 24362042

deVeaux86: de Veaux LC, Clevenson DS, Bradbeer C, Kadner RJ (1986). "Identification of the btuCED polypeptides and evidence for their role in vitamin B12 transport in Escherichia coli." J Bacteriol 1986;167(3);920-7. PMID: 3528128

Di11: Di Bartolo ND, Hvorup RN, Locher KP, Booth PJ (2011). "In vitro folding and assembly of the Escherichia coli ATP-binding cassette transporter, BtuCD." J Biol Chem 286(21);18807-15. PMID: 21345797

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Friedrich86: Friedrich MJ, de Veaux LC, Kadner RJ (1986). "Nucleotide sequence of the btuCED genes involved in vitamin B12 transport in Escherichia coli and homology with components of periplasmic-binding-protein-dependent transport systems." J Bacteriol 167(3);928-34. PMID: 3528129

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Goetz09: Goetz BA, Perozo E, Locher KP (2009). "Distinct gate conformations of the ABC transporter BtuCD revealed by electron spin resonance spectroscopy and chemical cross-linking." FEBS Lett 583(2);266-70. PMID: 19101549

Hvorup07: Hvorup RN, Goetz BA, Niederer M, Hollenstein K, Perozo E, Locher KP (2007). "Asymmetry in the structure of the ABC transporter-binding protein complex BtuCD-BtuF." Science 317(5843);1387-90. PMID: 17673622

Ivetac07: Ivetac A, Campbell JD, Sansom MS (2007). "Dynamics and function in a bacterial ABC transporter: simulation studies of the BtuCDF system and its components." Biochemistry 46(10);2767-78. PMID: 17302441

Joseph11: Joseph B, Jeschke G, Goetz BA, Locher KP, Bordignon E (2011). "Transmembrane gate movements in the type II ATP-binding cassette (ABC) importer BtuCD-F during nucleotide cycle." J Biol Chem 286(47);41008-17. PMID: 21953468

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Kandt10: Kandt C, Tieleman DP (2010). "Holo-BtuF stabilizes the open conformation of the vitamin B12 ABC transporter BtuCD." Proteins 78(3);738-53. PMID: 19847921

Kenley78: Kenley JS, Leighton M, Bradbeer C (1978). "Transport of vitamin B12 in Escherichia coli. Corrinoid specificity of the outer membrane receptor." J Biol Chem 253(5);1341-6. PMID: 342525

Korkhov12: Korkhov VM, Mireku SA, Hvorup RN, Locher KP (2012). "Asymmetric states of vitamin B₁₂ transporter BtuCD are not discriminated by its cognate substrate binding protein BtuF." FEBS Lett 586(7);972-6. PMID: 22569249

Korkhov12a: Korkhov VM, Mireku SA, Locher KP (2012). "Structure of AMP-PNP-bound vitamin B12 transporter BtuCD-F." Nature 490(7420);367-72. PMID: 23000901

Lewinson10: Lewinson O, Lee AT, Locher KP, Rees DC (2010). "A distinct mechanism for the ABC transporter BtuCD-BtuF revealed by the dynamics of complex formation." Nat Struct Mol Biol 17(3);332-8. PMID: 20173761

Locher02: Locher KP, Lee AT, Rees DC (2002). "The E. coli BtuCD structure: a framework for ABC transporter architecture and mechanism." Science 296(5570);1091-8. PMID: 12004122

Locher04: Locher KP, Borths E (2004). "ABC transporter architecture and mechanism: implications from the crystal structures of BtuCD and BtuF." FEBS Lett 564(3);264-8. PMID: 15111107

Oloo04: Oloo EO, Tieleman DP (2004). "Conformational transitions induced by the binding of MgATP to the vitamin B12 ATP-binding cassette (ABC) transporter BtuCD." J Biol Chem 279(43);45013-9. PMID: 15308647

Rioux89: Rioux CR, Kadner RJ (1989). "Vitamin B12 transport in Escherichia coli K12 does not require the btuE gene of the btuCED operon." Mol Gen Genet 1989;217(2-3);301-8. PMID: 2671656

Tamura12: Tamura M, Kers JA, Cohen SN (2012). "Second-site suppression of RNase E essentiality by mutation of the deaD RNA helicase in Escherichia coli." J Bacteriol 194(8);1919-26. PMID: 22328678

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

Wu95: Wu LF, Mandrand-Berthelot MA (1995). "A family of homologous substrate-binding proteins with a broad range of substrate specificity and dissimilar biological functions." Biochimie 1995;77(9);744-50. PMID: 8789466


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Sat Dec 20, 2014, BIOCYC14A.